Enterococcus faecalis is a member of the enterococci, a group of hardy organisms which have become the second leading cause of hospital-acquired infections in this country. E. faecalis is adept at transferring drug resistance to many of the antibiotics commonly used for treatment of enterococcal infections, such as aminoglycosides, penicillins, and glycopeptide antibiotics (e.g. vancomycin). The proportion of enterococci resistant to vancomycin has increased at an alarming rate in the last few years, and the frightening prospect looms of enterococci disseminating the high level vancomycin-resistance to pathogens, such as Staphylococcus aureus (for which vancomycin is often the last antimicrobial option). The therapeutic options for treatment of enterococcal infections are dwindling at a time when new classes of antibiotics are not on the horizon, threatening a return to the pre-antibiotic era. In an effort to rapidly provide the research community with new targets for antimicrobial therapy, the investigators propose to sequence the entire genome of a vancomycin-resistant strain of E. faecalis. Advances in high throughput sequencing and computer analysis have made this a feasible, cost-effective approach to achieve this goal. In the first phase of the project, approximately 46,000 forward and reverse sequence reads will be obtained from a library of small, randomly sheared fragments of the E. faecalis genome. The second phase of the project will consist of assembly of the random fragments into contiguous sequences (contigs) and closure of the gaps using methods developed and refined for the three other prokaryotic genome projects at TIGR. The final phase of the project will involve the annotation of the genome sequence to identify the salient features of the genome. For example, all coding regions will be identified and searched for homology against other known sequences, providing insight into the pathogenesis of enterococcal infections based on studies from other bacteria. Along with publication of the sequence analysis in a peer-reviewed journal, the finished product will be deposited into a variety of databases including the TIGR World Wide Web site. In addition, research materials such as clone sets will be made available to the research community. Upon completion of this project, a global picture of enterococcal biochemistry, genetics, and physiology will emerge and should allow the development of new antimicrobial therapies against this emerging pathogen.